Functional studies of the kidney of living animals using multicolor two-photon microscopy
Functional studies of the kidney of living animals using multicolor two-photon microscopy
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DOI：
10.1152/ajpcell.00159.2002
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发表时间：
2002-09-01
影响因子：
5.5
通讯作者：
Molitoris, BA
中科院分区：
文献类型：
--
作者：
Dunn, KW;Sandoval, RM;Molitoris, BA
当应用于活体动物时，光学显微镜为在最生理相关的环境中研究细胞生物学提供了强有力的手段。双光子显微镜能够收集生物组织深处的光学切片，这为活体显微镜领域开辟了一条道路，可以对大脑、透镜、皮肤和肿瘤进行高分辨率研究。在这里，我们提出的方式，双光子显微镜可以应用于肾脏生理学的活体研究的例子。因为肾脏很容易在不损害其功能的情况下被外部化，所以显微镜可用于评估体内肾功能的各个方面。这些包括细胞活力和凋亡，液体运输，受体介导的内吞作用，血流和白细胞运输。多个荧光团的有效双光子激发允许多个探针的比较和多个参数的同时表征，并产生光谱信息，这对于解释包含未表征的自体荧光的图像是至关重要的。这里描述的研究表明，双光子显微镜可以提供以前在活体显微镜无法达到的分辨率水平的方式，使动力学分析和生理学研究的活动物的器官与亚细胞分辨率。
Optical microscopy, when applied to living animals, provides a powerful means of studying cell biology in the most physiologically relevant setting. The ability of two-photon microscopy to collect optical sections deep into biological tissues has opened up the field of intravital microscopy to high-resolution studies of the brain, lens, skin, and tumors. Here we present examples of the way in which two-photon microscopy can be applied to intravital studies of kidney physiology. Because the kidney is easily externalized without compromising its function, microscopy can be used to evaluate various aspects of renal function in vivo. These include cell vitality and apoptosis, fluid transport, receptor-mediated endocytosis, blood flow, and leukocyte trafficking. Efficient two-photon excitation of multiple fluorophores permits comparison of multiple probes and simultaneous characterization of multiple parameters and yields spectral information that is crucial to the interpretation of images containing uncharacterized autofluorescence. The studies described here demonstrate the way in which two-photon microscopy can provide a level of resolution previously unattainable in intravital microscopy, enabling kinetic analyses and physiological studies of the organs of living animals with subcellular resolution.